Apoptosis induced by the Tibetan herbal remedy PADMA 28 in the T cell-derived lymphocytic leukaemia cell line CEM-C7H2

The Tibetan herbal remedy PADMA 28 revealed promising results to support treatment of atherosclerosis, Charot syndrome (intermittent claudication), chronic active hepatitis and infection of the respiratory tract. The remedy was confirmed to be closely linked with anti- and pro-oxidative properties in vitro. In this study, apoptogenic and survival effects of PADMA 28 were investigated in the T cell-derived lymphocytic leukaemia cell line CEM-C7H2. PADMA 28 led to a concentration-dependent inhibition of cell proliferation accompanied by the accumulation of CEM-C7H2 cells in subG1 phase, fragmentation of poly (ADP-ribose) polymerase (PARP) and nuclear body formation. Treatment with PADMA 28 rescued to some extent cells over-expressing Bcl-2 from apoptosis. This finding suggests that the mechanism of action of PADMA 28 may be via interference with Bcl-2 triggered survival pathways

Crinum Latifolium Leave Extracts Suppress Immune Activation Cascades in Peripheral Blood Mononuclear Cells and Proliferation of Prostate Tumor Cells

Plants of the genus Crinum (Amaryllidaceae) are widely used in folk medicine in different tropical and subtropical regions around the world. The Indian species Crinum latifolium (L.) was traditionally used to treat rheumatism, fistula, tumors, earaches, rubefacient, tubercle and whitlow. In Vietnamese and Chinese traditional medicine Crinum latifolium preparations are used until nowadays because of their antiviral and antitumor properties. In this study, we demonstrate potent in vitro antioxidant activity of an aqueous Crinum latifolium extract by an oxygen radical absorbance capacity (ORAC) value of 1610 ± 150 μmol Trolox equivalents/g. Furthermore, significant anti-inflammatory effects of this extract were shown by its potential to suppress indoleamine 2,3-dioxygenase (IDO) mediated tryptophan degradation in unstimulated- and mitogen-stimulated PBMC at IC50 doses of 241 ± 57 μg/ml and 92 ± 20 μg/ml, respectively. Concentrations of the immune activation marker neopterin were slightly diminished in unstimulated PBMC, whereas a dose-dependent inhibition of neopterin formation was observed in mitogen-stimulated PBMC (IC50 = 453 ± 86 μg/ml). Additionally, we measured also dose-dependent inhibitory effects of this aqueous Crinum latifolium extract on cell proliferation of highly metastatic human prostate carcinoma PC3 cells (IC50 = 4.5 ± 0.8 mg/ml), androgen-sensitive prostate adenocarcinoma LNCaP cells (IC50 =2.3 ± 0.1 mg/ml), and benign prostate hyperplasia BPH-1 cells (IC50 = 2.1 ± 0.04 mg/ml). We conclude that both effects, inhibition of tumor cell growth and recovery of immune functions, are important for the antitumor properties of Crinum latifolium

An update on the strategies in multicomponent activity monitoring within the phytopharmaceutical field

<p>Abstract</p> <p>Background</p> <p>To-date modern drug research has focused on the discovery and synthesis of single active substances. However, multicomponent preparations are gaining increasing importance in the phytopharmaceutical field by demonstrating beneficial properties with respect to efficacy and toxicity.</p> <p>Discussion</p> <p>In contrast to single drug combinations, a botanical multicomponent therapeutic possesses a complex repertoire of chemicals that belong to a variety of substance classes. This may explain the frequently observed pleiotropic bioactivity spectra of these compounds, which may also suggest that they possess novel therapeutic opportunities. Interestingly, considerable bioactivity properties are exhibited not only by remedies that contain high doses of phytochemicals with prominent pharmaceutical efficacy, but also preparations that lack a sole active principle component. Despite that each individual substance within these multicomponents has a low molar fraction, the therapeutic activity of these substances is established via a potentialization of their effects through combined and simultaneous attacks on multiple molecular targets. Although beneficial properties may emerge from such a broad range of perturbations on cellular machinery, validation and/or prediction of their activity profiles is accompanied with a variety of difficulties in generic risk-benefit assessments. Thus, it is recommended that a comprehensive strategy is implemented to cover the entirety of multicomponent-multitarget effects, so as to address the limitations of conventional approaches.</p> <p>Summary</p> <p>An integration of standard toxicological methods with selected pathway-focused bioassays and unbiased data acquisition strategies (such as gene expression analysis) would be advantageous in building an interaction network model to consider all of the effects, whether they were intended or adverse reactions.</p

Details of the behavioral experimental design and timeline.

<p>rmf: remifentanil, veh: vehicle, sal: saline, admin: administration, reacq: reaquisition, reinst: reinstatement, stau: staurosporine, zips: zip scramble, x: trials without drug reinforcement, ↓: intraaccumbens injection, rmf(1): self-administration of remifentanil only after completion of the first run.</p

Inhibition of AcbC PKC/Mzeta blocks memory consolidation.

<p>Rats were trained to traverse a runway alley to obtain a remifentanil (RMF; 0.032 mg/kg iv) injection paired with a light stimulus (conditioned stimulus, CS; blinking at 2 Hz for 20 s) on day A (acquisiton; top row). The PKCzeta- and PKMzeta-pseudosubstrate inhibitor ZIP (N = 7), scrambled ZIP peptide (N = 8), or the nonselective PKC inhibitor staurosporine (N = 7) was administered locally into the nucleus accumbens core (0.5microliter of 1.5 mM each) 30 min before runs #1 and #4. On day B, only the light stimulus (CS) was presented when the animal had traversed the runway (all groups). One day later (day C), all animals received i.v. RMF upon traversing the alley in run #1. Traversing the runway in run #2 had no consequences. The next day (day D), all animals were retrained with RMF plus the light CS. No RMF was administered in run #5 in order to determine goal times. Column 1, start time; column 2; alley time; column 3, goal time (not determined on day A because of direct sedative effects of RMF). The statistical analysis yielded the following results: Panel B1: 2W-RM-ANOVA: Interaction: F(6,19) = 1,0327; run number: F(3,19) = 40,51 p<0,0001; Group F(2,19) = 9,771; p<0,01, posthoc (Bonferroni): ZIP vs ZIP-scrambled: runs 1,3 p<0,05. ZIPscramble vs Staurosporine: NS. ZIP vs staurosporine, runs 2,3 p<0.05. Panel B3: 1W-ANOVA Only for 1^st run. F(2,19) = 24.870; p<0.001. Posthoc/Bonferroni: ZIP vs ZIP-scramble: p<0.001; ZIP vs Stauro: p<0.001; ZIP-scramble vs Stauro: NS. Panel C1: 1W-ANOVA Only for 2^nd run. F(2,19) = 9.341; p<0.001, posthoc (Bonferroni). ZIP vs ZIP-scramble d; p<0.01. ZIP vs STAURO, p<0.05. staurosporine vs ZIP-scrambled. NS. Panel C3: 1W-ANOVA: F(2,19) = 6.995; p<0.01. Posthoc (Bonferroni) ZIP vs ZIP-scrambled p<0.01. ZIP–scrambled vs staurosporine, NS. ZIP vs staurosporine, p<0.05. Panel D1: 2W-RM-ANOVA. Interaction: F(8,19) = 3,106 p<0.01; Run Number F(4,19) = 22,17 p<0.0001. Group: F(2,19) = 22.51 p<0.0001. Posthoc (Bonferroni): ZIP vs ZIP-scrambled: runs 1,2 (p<0.01) and 3 (p<0.05). ZIP-scrambled vs staurosporine. NS. ZIP vs staurosporine runs 1,2,3 p<0.01. Panel D3: 1-W-ANOVA: F(2,19) = 1.044. NS.</p

Local intra-AcbC inhibition of PKC/Mzeta the retrieval of conditioned remifentanil approach-associated memories.

<p>(A) Runway behavior before ZIP administration, runs #1-4: Drug-naïve male Sprague Dawley rats were given the opportunity to traverse a runway alley to obtain 0.032 mg/kg i.v. RMF (contingent RMF administration, i.e., RMF self-administration, 40 min inter-run interval; RMF, N = 13) or saline (SAL, N = 9) paired with a light stimulus (conditioned stimulus, CS; blinking at 2 Hz for 20 s)<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030502#pone.0030502-Crespo1" target="_blank">[8]</a> (B) Runway behavior after ZIP administration, run #5: In order to determine the goal time in absence of the drug with its confounding sedative effects, RMF was not administered in this last run. The PKCzeta- und PKMzeta-pseudosubstrate inhibitor ZIP (0.5microliter of 1.5 mM) was administered locally into the nucleus accumbens core (30 min before run #5) of five animals after the first four RMF runs (RMF+ZIP). Column 1 shows start time (i.e., latency to leave the start area), column 2 alley time (i.e., time needed to traverse the runway alley), and column 3 goal time (i.e., time spent in the goal area). In order to determine the effect of noncontingent RMF (i.e., the acute pharmacological effects of RMF) on PKCzeta- and PKMzeta activation (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030502#pone-0030502-g003" target="_blank">Fig. 3</a>), another group of animals passively received i.v. RMF within the confines of the runway (noncontingent RMF administration, N = 10). Their runway behavior was not recorded and is therefore not shown here. The statistical analysis gave the following results: Panel A1, 2W-RM-ANOVA, Interaction [F(6,19)] = 7,875 P<0.0001. Group [F(2,19)] = 16,44 P<0.0001, post-hoc (Bonferroni): ZIP vs Saline (run3: p<0.05, run4: p<0.001), RMF vs Saline(run3: p<0.001, run4:p<0.001). Panel B1: 1W-ANOVA F(2,19) = 52.90 p<0.0001, post-hoc (Bonferroni): RMF+ZIP vs Saline (p<0.001), RMF+ZIP vs RMF (ns), RMF vs Saline (p<0.001)). Panel B3, 1W-ANOVA F(2,19) = 13,66 p<0,0002, post-hoc (Bonferroni): RMF+ZIP vs Saline (ns), RMF+ZIP vs RMF (p<0.05), RMF vs Saline (p<0.001)).</p

Acquisition and expression of conditioned remifentanil approach is paralleled by an activaction of PKC/Mzeta in the AcbC.

<p>The animals, the behavior of which is shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030502#pone-0030502-g001" target="_blank">Fig. 1</a>, were sacrificed immediately after run #5 (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030502#pone-0030502-g001" target="_blank">Fig. 1B</a>), brain tissue lysates were obtained from the nucleus accumbens core (AcbC) and subcellular fractions were separated by ultracentrifugation. Ten micrograms of membrane- (m) or cytosolic (c) fraction were separated by SDS-PAGE, transferred to nitrocellulose membranes, and incubated with antibody (1∶1500) against PKCzeta (top panel, 100 kDa subunit band; middle panel; 75 kDa subunit band). This antibody also detected PKMzeta (bottom panel; 55 kDa band). Bound antibody was visualized with horseradish peroxidase-conjugated secondary antibody (1∶2000, Santa Cruz Biotechnology) and enhanced chemiluminescence. GAPDH immunoreactivity was used as a loading control. Activation of PKCzeta and PKMzeta is reflected by the membrane/cytosol ratio (abscissae; means ± SEM). S, saline (N = 9), NC, noncontingent remifentanil (N = 10), C, contingent remifentanil (N = 8), C+ZIP, contingent remifentanil followed by ZIP inhibition (N = 5). ANOVAs and Bonferroni post-hoc tests for each subunit gave the following results: 100 kDa subunit, F(3,28) = 3.001; p<0.05,* p<0.05 for C vs NC; 75 kDa subunit, F(3,28) = 3.698; p<0.023 * # p<0,05 for C vs NC and C vs S; and 55 kDa subunit, F(3,28) = 4,383; p<0.012; p<0.05 for C vs C+ZIP.</p